A nterior spinal pathological entities of the thoracolumbar spine have traditionally been treated by an anterior transthoracic, thoracolumbar, or retroperitoneal approach. Also used has been a posterior surgery via costotransversectomy or extracavitary approaches. The transpedicular corpectomy is a variant of the costotransversectomy and extracavitary approaches in that the rib head is preserved as the corpectomy is being performed, and the corpectomy is performed through the pedicles. Various techniques have been described to deal with the rib head to perform anterior spinal reconstruction. [4][5][6] With advances in surgical techniques, many operations that traditionally have been performed through anterior spinal approaches (including multilevel corpectomies) are now being performed via posterior approaches. We wished to evaluate whether one approach was superior to the other with regard to complications, neurological outcome, morbidity, and other perioperative factors. Object. Whereas standard anterior approaches for thoracolumbar corpectomies have commonly been used, the transpedicular technique is increasingly used to perform corpectomies from a posterior approach. The authors conducted a study to analyze whether there was a difference in outcomes by comparing transpedicular corpectomies to standard anterior thoracolumbar corpectomies. MethodsMethods. The senior author performed thoracolumbar corpectomies in 80 patients between 2004 and 2008. The authors reviewed medical records and follow-up data, consisting of clinic visits, radiographs, or telephone interviews. Neurological outcome, complications, operative times, revision surgery rates, and estimated blood loss (EBL) were evaluated.Results. Thirty-four patients underwent transpedicular corpectomies, and 46 patients underwent anterior thoracolumbar approaches. Single-level transpedicular corpectomies appear to be comparable to anterior-only corpectomies in terms of EBL, operative time, and complication rates. There was a higher complication rate, increased EBL, and longer operative time with anterior-posterior corpectomies compared with transpedicular corpectomies. Patients undergoing transpedicular corpectomies had a greater recovery of neurological function than those in whom anteriorapproach corpectomies were performed.Conclusions. The transpedicular corpectomy appears to have a comparable morbidity rate to anterior-only corpectomies, but its morbidity rate is lower than that of anterior-posterior corpectomies.

Combination treatment for non–small cell lung cancer (NSCLC) is becoming more popular due to the anticipation that it may be more effective than single drug treatment. In addition, there are efforts to genetically screen patients for specific mutations in light of attempting to administer specific anticancer agents that are most effective. In this study, we evaluate the anticancer and anti-angiogenic effects of low dose erlotinib-cisplatin combination in NSCLC in vitro and in vivo. In NSCLC cells harboring epidermal growth factor receptor (EGFR) mutations, combination erlotinib-cisplatin treatment led to synergistic cell death, but there was minimal efficacy in NSCLC cells with wild-type EGFR. In xenograft models, combination treatment also demonstrated greater inhibition of tumor growth compared to individual treatment. The anti-tumor effect observed was secondary to the targeting of angiogenesis, evidenced by decreased vascular endothelial growth factor (VEGF) levels and decreased levels of CD31 and microvessel density. Combination treatment targets angiogenesis through down-regulation of the c-MYC/hypoxia inducible factor 1-alpha (HIF-1α) pathway. In fact, cell lines with EGFR exon 19 deletions expressed high basal levels of c-MYC and HIF-1α and correlate with robust responses to combination treatment. These results suggest that low dose erlotinib-cisplatin combination exhibits its anti-tumor activity by targeting angiogenesis through the modulation of the c-MYC/HIF-1α/VEGF pathway in NSCLC with EGFR exon 19 deletions. These findings may have significant clinical implications in patients with tumors harboring EGFR exon 19 deletions as they may be particularly sensitive to this regimen.

Tyrosine kinase inhibitors such as erlotinib are commonly used as a therapeutic agent against cancer due to its relatively low side-effect profile and, at times, greater efficacy. However, erlotinib resistance (ER) in non-small cell lung cancer is being recognized as a major problem. Therefore, understanding the mechanism behind ER and developing effective regimens are needed. Autophagy’s role in cancer has been controversial and remains unclear. In this study, we examined the effectiveness of low dose erlotinib-cisplatin combination in erlotinib resistant lung adenocarcinoma (ERPC9) cells and the role of autophagy in ER. ERPC9 cells were established from erlotinib sensitive PC9 cells. Appropriate treatments were done over two days and cell survival was quantified with Alamar Blue assay. LC3II and regulatory proteins of autophagy were measured by western blot. Small interfering RNA (siRNA) was utilized to inhibit translation of the protein of interest. In ERPC9 cells, combination treatment induced synergistic cell death and a significant decrease in autophagy. At baseline, ERPC9 cells had a significantly higher LC3II and lower p-mTOR levels compared to PC9 cells. The addition of rapamycin increased resistance and 3-methyladenine sensitized ERPC9 cells, indicating autophagy may be acting as a protective mechanism. Further examination revealed that ERPC9 cells harbored high baseline Atg3 levels. The high basal Atg3 was targeted and significantly lowered with combination treatment. siRNA transfection of Atg3 resulted in the reversal of ER; 42.0% more cells died in erlotinib-alone treatment with transfection compared to non-transfected ERPC9 cells. We reveal a novel role for Atg3 in the promotion of ER as the inhibition of Atg3 translation was able to result in the re-sensitization of ERPC9 cells to erlotinib-alone treatment. Also, we demonstrate that combination erlotinib-cisplatin is an effective treatment against erlotinib resistant cancer by targeting (down-regulating) Atg3 mediated autophagy and induction of apoptotic cell death.

Previous studies of word segmentation in a second language have yielded equivocal results. This is not surprising given the differences in the bilingual experience and proficiency of the participants and the varied experimental designs that have been used. The present study tried to account for a number of relevant variables to determine if bilingual listeners are able to use native-like word segmentation strategies. Here, 61 French-English bilingual adults who varied in L1 (French or English) and language dominance took part in an audiovisual integration task while event-related brain potentials (ERPs) were recorded. Participants listened to sentences built around ambiguous syllable strings (which could be disambiguated based on different word segmentation patterns), during which an illustration was presented on screen. Participants were asked to determine if the illustration was related to the heard utterance or not. Each participant listened to both English and French utterances, providing segmentation patterns that included both their native language (used as reference) and their L2. Interestingly, different patterns of results were observed in the event-related potentials (online) and behavioral (offline) results, suggesting that L2 participants showed signs of being able to adapt their segmentation strategies to the specifics of the L2 (online ERP results), but that the extent of the adaptation varied as a function of listeners' language experience (offline behavioral results).

Introduction: Non-small cell lung cancer (NSCLC) accounts for about 80% of all cases of lung cancer. The treatment of choice for NSCLC is complete surgical resection. However, in advance stages with metastasis and/or aggressive local infiltrate chemotherapy becomes the only option. At high therapeutic doses, chemotherapy can cause significant side effects, and the development of resistance may arise if initial treatment doses of therapy become ineffective. In order to prevent serious side effects and resistance, low-dose combination regimens that offer comparable results to current high-dose regimens are needed. The combination of cisplatin and erlotinib has been shown to inhibit tumor growth more effectively than a single drug in NSCLC cells. However, the mechanism underlying this finding remains unclear. Autophagy has been suggested to play a critical role in cancer and chemotherapy, but there is ongoing controversy among its role (promotion vs. prevention of cancer). In this study, we examined whether the combination of low-dose cisplatin and erlotinib is able to induce higher levels of cancer cell death than each drug alone in both sensitive and erlotinib-resistant lung adenocarcinoma. In addition, we looked into the role of autophagy in cancer cells and its association with chemotherapy. Methods: Alamar blue assay was used to construct dose response curves in which IC 50 values of cisplatin and erlotinib were determined. Erlotinib-resistant PC9 (PC9/ER) cells were developed by culturing PC9 (EGFR overexpression adenocarcinoma) cells in media containing an erlotinib dose 1000 times the IC50 value over a two week period; PC9/ER cells were verified to be 26-fold more resistant than PC9 sensitive cancer cells. Baseline levels of autophagy of normal human bronchial epithelial (NHBE) cells, PC9, and PC9/ER were examined by Western blot with antibodies against LC3I/II. The IC 25 of each drug was chosen as the low-dose treatments (3 uM cisplatin and 10 nM erlotinib). Cells were treated accordingly: control, cisplatin, erlotinib, and combination. Cell death and autophagy levels were measured. Next, cells were pretreated in one day in advance with rapamycin and cell death and changes in autophagy were assessed. Results: At baseline, there were higher levels of LC3II in PC9/ER compared to PC9 and NHBE. Combined treatment of cisplatin and erlotinib was able to induce significantly more cell death than individual drugs at the same dose (p<0.001). In fact, combination treatment provided synergistic killing effects in PC9 and PC9/ER cells. In PC9 cells, erlotinib alone had 79.9% survival, cisplatin 76.3%, and combination 45.0%. In PC9/ER, erlotinib alone had 96.6% survival, cisplatin 89.3%, and combination 61.1%. Levels of autophagy decreased in a similar pattern to killing-effects in both PC9 and PC9/ER cells. In addition, a significant increase in cell survival of about 20% was observed when pretreated with rapamycin (p<0.001). NHBE demonstrated no significant killing and no changes in autophagy levels. Conclusion: Our data is consistent for autophagy's role in promoting cancer cell survival, work as a protective mechanism for cancer-resistant cells, and a critical mechanism in chemotherapy-induced cancer cell death. The combination of erlotinib and cisplatin offers synergistic killing properties in vitro and may be a potential regimen against resistant lung cancer. In addition, the combination treatment does not induce significant cytotoxic effects and death in NHBE cells indicating its safe dosing. We plan to further characterize autophagy with 3-methyladenine pretreatment, changes in apoptosis, and invasiveness of cancer cells. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the Second AACR International Conference on Frontiers in Basic Cancer Research; 2011 Sep 14-18; San Francisco, CA. Philadelphia (PA): AACR; Cancer Res 2011;71(18 Suppl):Abstract nr C59.

Objective Erlotinib resistance (ER) in non‐small cell lung cancer (NSCLC) is a major problem clinically. Therefore, in this study we tested whether combination of low dose erlotinib and cisplatin is effective against ER and specifically examined the role of autophagy in ER. Methods PC9 cells were used to establish erlotinib resistant cells (PC9ER). Cell survival was quantified with Alamar blue assay. LC3II and regulatory proteins of autophagy were measured by western blot. SiRNA was used to modulate protein translation. Results Combination of low dose erlotinib‐cisplatin induced synergistic cell death in PC9ER cells and this response was related to a decrease in autophagy. Concordantly, rapamycin increased ER and 3‐MA sensitized cell death response in PC9ER cells. Western blot analysis of autophagy regulatory proteins showed significantly higher baseline levels of LC3II and Atg3 in ER cells as compared to the sensitive PC9 and a significant decrease in Atg3 level was seen with combination treatment. Atg3 SiRNA transfection resulted in the reversal of the resistance in PC9ER cells to both erlotinib‐alone and combination treatment. Conclusions Our data suggests that ER lung cancer cells are responsive to combined low dose erlotinib‐cisplatin treatment by targeting the baseline over production of Atg3 and that Atg3 is a key regulator/promoter of ER in NSCLC. This research was supported by NIH‐T32 grant.

We have recently reported elevated MARCKS phosphorylation (p-MARCKS) in lung cancer tissues but not in their adjacent normal non-cancer tissue sections. We have extended this study to a tumor tissue array from a cohort of 110 human patients with lung cancer. Immunohistochemistry results have shown a significant association of elevated p-MARCKS with advanced-stage, lymph node metastasis and malignant phenotypes of lung cancer, confirming the importance of p-MARCKS in lung cancer progression. To see if this newly discover biomarker can be a target for therapeutic treatment for the suppression of lung cancer malignancy, we have employed two peptides for this potential; one is the MANS peptide developed by our collaborator many years ago to target myristoylated N-terminal amino acid region for the control of airway mucus granule secretion, the other peptide is MPS that directly targets the MARCKS phosphorylation site domain. For MANS peptide, our recent publication has shown it is effective in blocking lung cancer cell migration and invasiveness in vitro as well as in vivo metastasis. A similar observation was seen for MPS peptide treatment. However, in contrast to MANS peptide, MPS treatment can directly block PIP3-dependent AKT signaling and repress lung tumor growth in vivo in addition to the suppression of metastasis, while MANS can only reduce cancer metastasis. These results support the initial notion on the involvement of p-MARCKS as a biomarker in lung cancer and the potential to target the phosphorylation site domain for controlling cancer progression. Our data have also suggested a direct targeting MARCKS phosphorylation site domain by MPS peptide is more effective in the suppression of lung cancer malignancy. Citation Format: Ching-Hsien Chen, Sarah Statt, Phillip Thai, Jasmine G. Lee, Kenneth B. Adler, Pan-Chyr Yang, Reen Wu. Suppression of lung cancer malignancy by peptides targeting MARCKS phosphorylation site domain: A comparative study. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2596. doi:10.1158/1538-7445.AM2014-2596

Introduction: Platinum based agents such as cisplatin have been traditionally used to for the treatment of lung cancer, and tyrosine kinase inhibitors such as erlotinib are being used more readily. Combinations of anti-cancer agents are becoming more popular due to the anticipation that combination treatment is more effective than individual drug treatments. In this study, we evaluate the anti-cancer effects of combination erlotinib and cisplatin in in vitro cell lines and in in vivo mice models. In addition, as studies of cisplatin and erlotinib in other cancer types show potential to inhibit angiogenesis, we examine whether these agents are able to target angiogenesis in lung adenocarcinoma. Method: Three individual lung adenocarcinoma cell lines (PC9, A549, and H292) were utilized. Each cell line had four treatment groups: control, erlotinib only, cisplatin only, and combination erlotinib and cisplatin. Alamar Blue Assay was utilized to evaluate for in vitro cell death following two day treatments. PC9 cells were injected into the left flank of nude mice for in vivo experiments. Erlotinib and/or cisplatin treatments were injected directly into the tumor and tumor measurements were made every 2 days for a total of 14 days. Western blot analysis of VEGFR, p-VEGFR, and CD31was performed for both in vitro and in vivo samples. Hematoxylin and eosin (H&E) staining and 3,3′-Diaminobenzidine (DAB) staining for CD31 were done on tumor samples. Results: Combination treatment in EGFR mutated PC9 cells led to significantly higher cell death compared to other treatment groups (P<0.0001); the effect was synergistic. In EGFR wild type A549 and H292 cell lines there were no significant differences. Similar to results from cell cultures, in vivo studies of combination treatment showed significant inhibition of tumor growth compared to other treatment groups (p=0.001). On western blot analysis, there were significantly lower levels of p-VEGFR (p<0.0001) and CD31 (p=0.0291) in the combination group compared to all other groups. H&E staining shows significantly lower density of blood vessel and DAB staining of CD31in the combination group compared to other groups. Conclusions: Combination low dose erlotinib and cisplatin is able to induce synergistic cell death. This effect, however, may be specific to EGFR mutated lung adenocarcinoma. Combination treatment also demonstrated the ability to inhibit tumor growth in mice. The inhibition of tumor growth may be secondary to inhibition of angiogenesis. Citation Format: Jasmine G. Lee, Reen Wu. Combination erlotinib and cisplatin causes decrease angiogenesis in lung adenocarcinoma. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2068. doi:10.1158/1538-7445.AM2013-2068